Purified alpha subunits from human chorionic gonadotropin, TSH, FSH, and LH will be treated with neuraminidase and then subjected to alkaline borohydride degradation followed by trifluoroacetolysis. Oligosaccharides released by the alkaline borohydride step will be studied by gel filtration, methylation analysis and mass spectrometry of the permethylated oligosaccharide derivatives. Conditions for trifluoroacetolysis will be adjusted so as to destroy reducing amino sugars after release of oligosaccharides from chitobiosyl-asparagine linkages. Following removal of N-trifluoroacetyl groups from any remaining amino sugars in the mixture, oligosaccharides will be subjected to ion exchange chromatography to separate "high mannose" from "complex" type chains. The oligosaccharides obtained will be subjected to gel filtration chromatography, high voltage electrophoresis in borate buffer, and paper chromatography to investigate possible heterogeneity of carbohydrate chains. Fractions will be monitored by sugar analysis at each step.